Stevia composition

ABSTRACT

Stevia compositions are prepared from steviol glycosides of  Stevia rebaudiana  Bertoni. The compositions are able to provide a superior taste profile and can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals.

This application is a continuation of application Ser. No. 16/036,440, filed on Jul. 16, 2018, which is a continuation of application Ser. No. 14/118,673, filed on Feb. 4, 2014, now U.S. Pat. No. 10,021,899, which is a § 371 national stage entry of PCT Application No. PCT/US2011/047498, filed on Aug. 11, 2011, which claims priority under 35 U.S.C. § 119(e) from provisional Patent Application No. 61/491,441, filed on May 31, 2011, the contents of which applications are incorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The invention relates to a process for producing a highly purified food ingredient from the extract of the Stevia rebaudiana Bertoni plant and its use in various food products and beverages.

Description of the Related Art

Sugar alternatives are receiving increasing attention due to the awareness of many diseases associated with the consumption of high-sugar foods and beverages. However, many artificial sweeteners such as dulcin, sodium cyclamate and saccharin have been banned or restricted in some countries due to concerns about their safety. As a result, non-caloric sweeteners of natural origin are becoming increasingly popular. The sweet herb Stevia rebaudiana Bertoni produces a number of diterpene glycosides which feature high intensity sweetness and sensory properties superior to those of many other high potency sweeteners.

The above-mentioned sweet glycosides, have a common aglycon, steviol, and differ by the number and type of carbohydrate residues at the C13 and C19 positions. The leaves of Stevia are able to accumulate up to 10-20% (on dry weight basis) steviol glycosides. The major glycosides found in Stevia leaves are rebaudioside A (2-10%), stevioside (2-10%), and rebaudioside C (1-2%). Other glycosides such as rebaudioside B, D, E, and F, steviolbioside and rubusoside are found at much lower levels (approx. 0-0.2%).

Two major glycosides—stevioside and rebaudioside A (reb A), were extensively studied and characterized in terms of their suitability as commercial high intensity sweeteners. Stability studies in carbonated beverages confirmed their heat and pH stability (Chang S. S., Cook, J. M. (1983) Stability studies of stevioside and rebaudioside A in carbonated beverages. J Agric. Food Chem. 31: 409-412.)

Steviol glycosides differ from each other not only in their molecular structures, but also by their taste properties. Usually stevioside is found to be 110-270 times sweeter than sucrose, rebaudioside A between 150 and 320 times sweeter than sucrose, and rebaudioside C between 40-60 times sweeter than sucrose. Dulcoside A is 30 times sweeter than sucrose. Rebaudioside A has the least astringent, the least bitter, and the least persistent aftertaste, thus possessing the most favorable sensory attributes in major steviol glycosides (Tanaka O. (1987) Improvement of taste of natural sweetners. Pure Appl. Chem. 69:675-683; Phillips K. C. (1989) Stevia: steps in developing a new sweetener. In: Grenby T. H. ed. Developments in sweeteners, vol. 3. Elsevier Applied Science, London. 1-43.) The chemical structure of rebaudioside A is shown in FIG. 1 .

Methods for the extraction and purification of sweet glycosides from the Stevia rebaudiana plant using water or organic solvents are described in, for example, U.S. Pat. Nos. 4,361,697; 4,082,858; 4,892,938; 5,972,120; 5,962,678; 7,838,044 and 7,862,845.

However, even in a highly purified state, steviol glycosides still possess undesirable taste attributes such as bitterness, sweet aftertaste, licorice flavor, etc. One of the main obstacles for the successful commercialization of stevia sweeteners are these undesirable taste attributes. It was shown that these flavor notes become more prominent as the concentration of steviol glycosides increases (Prakash I., DuBois G E., Clos J. F., Wilkens K. L., Fosdick L. E. (2008) Development of rebiana, a natural, non-caloric sweetener. Food Chem. Toxicol., 46, S75-S82).

Rebaudioside B (CAS No: 58543-17-2), or reb B, also known as stevioside A₄ (Kennelly E. J. (2002) Constituents of Stevia rebaudiana In Stevia: The genus Stevia, Kinghorn A. D. (Ed), Taylor & Francis, London, p. 71), is one of the sweet glycosides found in Stevia rebaudiana. Sensory evaluations show that reb B was approximately 300-350 times sweeter than sucrose, while for reb A this value was approximately 350-450 (Crammer, B. and Ikan, R. (1986) Sweet glycosides from the Stevia plant. Chemistry in Britain 22, 915-916, and 918). The chemical structure of rebaudioside B is shown in FIG. 2 .

It was believed that reb B forms from the partial hydrolysis of rebaudioside A during the extraction process (Kobayashi, M., Horikawa, S., Degrandi, I. H., Ueno, J. and Mitsuhashi, H. (1977) Dulcosides A and B, new diterpene glycosides from Stevia rebaudiana. Phytochemistry 16, 1405-1408). However, further research has shown that reb B occurs naturally in the leaves of Stevia rebaudiana and is currently one of nine steviol glycosides recognized by FAO/JECFA (United Nations' Food and Agriculture Organization/Joint Expert Committee on Food Additives) in calculating total steviol glycosides' content in commercial steviol glycoside preparations (FAO JECFA (2010) Steviol Glycosides, Compendium of Food Additive Specifications, FAO JECFA Monographs 10, 17-21).

Only a few methods are described in literature for preparing reb B.

Kohda et al., (1976) prepared reb B by hydrolysis of reb A with hesperidinase. Reb B was also prepared by alkaline saponification of reb A. The said saponification was conducted in 10% potassium hydroxide-ethanol. The solution was acidified with acetic acid, and extracted with n-butanol. The butanol layer was washed with water and concentrated at low temperature in vacuo. The residue was crystallized from methanol to give reb B. (Kohda, H., Kasai, R., Yamasaki, K., Murakami, K. and Tanaka, O. (1976) New sweet diterpene glucosides from Stevia rebaudiana. Phytochemistry 15, 981-983). The described processes might be suitable for laboratory scale preparation of reb B, but are not suitable for any large scale or commercial reb B preparation.

Ahmed et al., used mild alkaline hydrolysis of reb A to prepare reb B. According to the described procedure, reb A was hydrolyzed to reb B by refluxing with 10% aqueous KOH at 100° C. for 1 hr. After neutralization with glacial acetic acid, the precipitated substance was recrystallized twice from methanol (Ahmed M. S., Dobberstein R. H., and Farnsworth N. R. (1980) Stevia rebaudiana: I. Use of p-bromophenacyl bromide to enhance ultraviolet detection of water-soluble organic acids (steviolbioside and rebaudioside B) in high-performance liquid chromatographic analysis, J. Chromatogr., 192, 387-393).

The use of methanol as recrystallization media as described in the literature will require its subsequent removal from the product. It is noted that handling of toxic substances such as methanol requires specialized manufacturing installations and, when applied in food processing, sophisticated food safety measures.

It is also noted that no significant work has been conducted to determine the potential of reb B as a sweetener or food ingredient. Moreover, reb B is often viewed as process artifact and unnecessary impurity in commercial steviol glycosides preparations. No significant evaluation of the influence of reb B on the overall taste profile of steviol glycoside preparations has been conducted.

The water solubility of reb B is reported to be about 0.1% (Kinghorn A. D. (2002) Constituents of Stevia rebaudiana In Stevia: The genus Stevia, Kinghorn A. D. (Ed), Taylor & Francis, London, p. 8). In many food processes where highly concentrated ingredients are used, a highly soluble form of reb B will be necessary.

Considering the facts mentioned above, there is a need to evaluate reb B as a sweetener and food ingredient and to develop a simple and efficient process for food grade reb B preparations suitable for food and other applications.

It is further noted that by having a carboxyl group in the molecule makes it possible for reb B to exist in the forms of various carboxylate salts. Prior to this invention, it is not believed that different carboxylate salt form of reb B had been prepared or evaluated for their impact on taste profiles.

Within the description of this invention we will show that, when applied in specific manner, carboxylate salts of reb B may impact the taste profile and offer significant advantages for the use of stevia sweeteners in various applications.

SUMMARY OF THE INVENTION

The present invention is aimed to overcome the disadvantages of existing Stevia sweeteners. The invention describes a process for producing a high purity food ingredient from the extract of the Stevia rebaudiana Bertoni plant and use thereof in various food products and beverages as a sweetness and flavor modifier.

The invention, in part, pertains to an ingredient comprising steviol glycosides of Stevia rebaudiana Bertoni plant. The steviol glycodsides are selected from the group consisting of stevioside, rebaudioside A (FIG. 1 ), rebaudioside B (FIG. 2 ), rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, steviolbioside, rubusoside, as well as other steviol glycosides found in Stevia rebaudiana Bertoni plant and mixtures thereof.

The invention, in part, pertains to a process for producing an ingredient containing rebaudioside B, and stevioside, rebaudioside A, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, steviolbioside, rubusoside, as well as other steviol glycosides found in Stevia rebaudiana Bertoni plant and mixtures thereof.

In the invention, rebaudioside A commercialized by PureCircle Sdn. Bhd. (Malaysia), containing, rebaudioside A (about 95-100%), stevioside (about 0-1%), rebaudioside C (about 0-1%), rebaudioside F (about 0-1%), rebaudioside B (about 0.1-0.8%), rebaudioside D (about 0-1%), and other glycosides amounting to total steviol glycosides' content of at least 95%, may be used as a starting material. Alternatively stevia extracts with different ratios of steviol glycosides may be used as starting materials.

In one embodiment, the starting material is subjected to partial or full conversion into reb B using an alkaline conversion process. The obtained reb B is then transformed into a carboxylate salt form by interacting with the respective base. These salt forms are utilized in similar manner as “original” reb B with a carboxyl group.

The obtained reb B or its carboxylate salt forms, and mixtures thereof may be subjected to additional thermal treatment to increase solubility.

The obtained products were applied in various foods and beverages as sweeteners, sweetener enhancers and flavor modifiers, including soft drinks, ice cream, cookies, bread, fruit juices, milk products, baked goods and confectionary products.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention as claimed.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings are included to provide a further understanding of the invention. The drawings illustrate embodiments of the invention and together with the description serve to explain the principles of the embodiments of the invention.

FIG. 1 shows the chemical structure of rebaudioside A.

FIG. 2 shows the chemical structure of rebaudioside B

FIG. 3 shows an HPLC chromatogram of a stevia composition comprising rebaudioside A and rebaudioside B.

DETAILED DESCRIPTION OF THE INVENTION

Advantages of the present invention will become more apparent from the detailed description given hereinafter. However, it should be understood that the detailed description and specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

Rebaudioside A commercialized by PureCircle Sdn. Bhd. (Malaysia), containing, rebaudioside A (about 95-100%), stevioside (about 0-1%), rebaudioside C (about 0-1%), rebaudioside D (about 0-1%), rebaudioside F (about 0-1%), rebaudioside B (about 0.1-0.8%) and other glycosides amounting to total steviol glycosides' content of at least about 95%, may be used as a starting material. Alternatively stevia extracts with different ratios of steviol glycosides may be used as starting materials.

The HPLC analysis of the raw materials and products can be performed on an Agilent Technologies 1200 Series (USA) liquid chromatograph, equipped with Phenomenex Prodigy ODS3, 5 (4.6×250 mm) column at 40° C. The mobile phase was 32:68 mixture of acetonitrile and 10 mmol/L sodium phosphate buffer (about pH 2.6) at 1 mL/min. A diode array detector set at 210 nm can be used as the detector. One example of an HPLC chromatogram thus obtained is shown in FIG. 3 .

As used herein, unless specified further, “reb B” and “reb B composition” shall be used interchangeably to refer to purified rebaudioside B or rebaudioside B in combination with any other chemical entity.

Preparation of Reb B

1. Alkaline Conversion

One way to obtain reb B starting with reb A is described as follows. Reb A is dispersed in aqueous alkaline solution. The concentration of reb A is about 0-50% (w/v) preferably about 10-25%. The preferred alkaline agents include potassium hydroxide and sodium hydroxide, however other agents capable of increasing the pH of the media above about pH 7 may be used as well or alternatively. The concentration of alkaline agents is about 0.05-2.0M, preferably about 0.1-1.0M. The mixture is incubated at about 10-150° C., preferably about 30-100° C., for a period of about 0.5-48 hrs, preferably about 1-24 hrs. As a result, reb A is hydrolyzed to reb B. The molar yield of conversion of reb B is about 5-100%, preferably about 10-90%.

After the reaction, the alkaline agent is neutralized by an acid, preferably by sulfuric acid or ortho-phosphoric acid until a pH of about 3.0-5.0 is reached, preferably until a pH of about 3.0-4.0 is reached. Upon neutralization, a precipitate is formed. The precipitate is separated by any method known in the art such as filtration or centrifugation and washed with water until the water reaches a pH of about 4.0-5.0. The obtained crystalline material is dried under vacuum at about 60-105° C. to yield a mixture of reb A and reb B having a ratio of about 5%:95% to about 95%:5% (w/w), preferably about 50%:50% to about 90%:10% (w/w).

2. Optional Post-Conversion Purification

To obtain purified reb B, in one embodiment the separated precipitate described above is suspended in water and the mixture is subjected to continuous agitation over about 0.5-24 hrs, preferably about 1-3 hours, at about 50-100° C., preferably about 60-80° C. The ratio of precipitate to water (w/v) is about 1:5 to about 1:20, preferably about 1:10 to about 1:15. The washed crystals are separated and dried under vacuum at about 60-105° C. to yield reb B with about 99% purity.

3. Optional Post-Conversion Solubility Enhancement

The following procedure can be used to increase the water solubility of reb B or any reb B composition. The obtained compositions generally have a water solubility of less than about 0.2% (w/v). In order to increase the solubility of these compositions, the reb B composition is combined with the water at ratio of about 1:1 (w/w) and the obtained mixture is subjected to a gradient heat treatment which results in a high stability and high concentration solution. The gradient of about 1° C. per minute is used in heating the mixture. The mixture is heated to the temperature of about 110-140° C., preferably about 118-125° C. and is held at maximum temperature for about 0-120 min, preferably about 50-70 min. After the heat treatment, the solution is cooled down to room temperature at gradient of about 1° C. per minute. The solution is spray dried by a laboratory spray drier operating at about 175° C. inlet and about 100° C. outlet temperatures. An amorphous form of the composition is obtained with greater than about 20% solubility in water at room temperature.

Preparation of Reb B Salt

Reb B obtained using the processes described above, or any other process, can be fully or partially converted into a carboxylate salt form. Reb B, or a composition containing reb B, preferably reb B with a purity greater than 90% (w/w), is dispersed in the water to make an aqueous dispersion with 5-50% (w/v), preferably 5-15% solids content. Excess base is added to achieve a pH level of 6.5-14.0, preferably 8.5-11.0. The obtained mixture is incubated for 0.1-24 hrs, preferably 1-3 hrs. Then the suspended solids are separated by filtration and washed with water until neutral pH of the washing water is achieved. Alternatively other reactions able to convert reb B into a carboxylate salt form may be used. The preferred cations are K⁺ and Na⁺, and the respective bases—KOH and NaOH—are used. However other carboxylate salts of reb B can be prepared in a similar manner by using the base corresponding to the desired carboxylate salt.

The obtained reb B carboxylate salt can be further purified and its solubility enhanced as described above.

While not intending to be bound by theory, it is believed that any steviol glycoside having a carboxyl group can be converted into its carboxylate salt form for improving the taste and/or sweetness profile of the molecule. Another example of a steviol glycoside containing a carboxyl group is steviolbioside.

Use of Reb B Carboxylate Salt

The reb B carboxylate salt described above can be used as a sweetness enhancer, a flavor enhancer and/or a sweetener in various food and beverage products. Non-limiting examples of food and beverage products include carbonated soft drinks, ready to drink beverages, energy drinks, isotonic drinks, low-calorie drinks, zero-calorie drinks, sports drinks, teas, fruit and vegetable juices, juice drinks, dairy drinks, yoghurt drinks, alcohol beverages, powdered beverages, bakery products, cookies, biscuits, baking mixes, cereals, confectioneries, candies, toffees, chewing gum, dairy products, flavored milk, yoghurts, flavored yoghurts, cultured milk, soy sauce and other soy base products, salad dressings, mayonnaise, vinegar, frozen-desserts, meat products, fish-meat products, bottled and canned foods, tabletop sweeteners, fruits and vegetables.

Additionally the reb B carboxylate salt compositions can be used in drug or pharmaceutical preparations and cosmetics, including but not limited to toothpaste, mouthwash, cough syrup, chewable tablets, lozenges, vitamin preparations, and the like.

The compositions can be used “as-is” or in combination with other sweeteners, flavors and food ingredients.

Non-limiting examples of sweeteners include steviol glycosides, stevioside, rebaudioside A, rebaudioside B, rebaudioside C, rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, steviolbioside, rubusoside, as well as other steviol glycosides found in Stevia rebaudiana Bertoni plant and mixtures thereof, stevia extract, Luo Han Guo extract, mogrosides, high-fructose corn syrup, corn syrup, invert sugar, fructooligosaccharides, inulin, inulooligosaccharides, coupling sugar, maltooligosaccharides, maltodextins, corn syrup solids, glucose, maltose, sucrose, lactose, aspartame, saccharin, sucralose, sugar alcohols.

Non-limiting examples of flavors include lemon, orange, fruit, banana, grape, pear, pineapple, bitter almond, cola, cinnamon, sugar, cotton candy, vanilla flavors.

Non-limiting examples of other food ingredients include flavors, acidulants, organic and amino acids, coloring agents, bulking agents, modified starches, gums, texturizers, preservatives, antioxidants, emulsifiers, stabilisers, thickeners, gelling agents.

The following examples illustrate various embodiments of the invention. It will be understood that the invention is not limited to the materials, proportions, conditions and procedures set forth in the examples, which are only illustrative.

Example 1 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia), containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C, 0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D was dispersed in 1000 mL aqueous KOH (1M) and incubated at 50° C. for 2 hours. The mixture temperature was decreased to 20° C. and the pH was adjusted to pH 4.0 with sulfuric acid. The solution was held under moderate agitation conditions for 4 hours and a precipitate was formed. The precipitate was filtered and washed on the filter with 2000 mL of water. The washed crystals were dried under vacuum to yield 86 g material containing about 84% reb A and 16% reb B. The water solubility (at 25° C.) of obtained material was about 0.2% (w/v).

Example 2 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia), containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C, 0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D was dispersed in 1000 mL aqueous KOH (1M) and incubated at 80° C. for 5 hours. The mixture temperature was decreased to 20° C. and the pH was adjusted to about pH 4.0 with sulfuric acid. The solution was held under moderate agitation conditions for 4 hours and a precipitate was formed. The precipitate was filtered and washed on the filter with 2000 mL of water. The washed crystals were dried under vacuum to yield about 75 g material containing about 9% reb A and about 91% reb B. The water solubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 3 Preparation of Stevia Composition

100 g of rebaudioside A produced by PureCircle Sdn. Bhd. (Malaysia), containing, 98.1% rebaudioside A, 0.3% stevioside, 0.2 rebaudioside C, 0.2% rebaudioside F, 0.4% rebaudioside B and 0.6% rebaudioside D was dispersed in 1000 mL aqueous KOH (1M) and incubated at 80° C. for 7 hours. The mixture temperature was decreased to 20° C. and the pH was adjusted to about pH 4.0 with sulfuric acid. The solution was held under moderate agitation conditions for 3-4 hours and a precipitate was formed. The precipitate was filtered and washed on the filter with 2000 mL of water. The washed crystals were dried under vacuum to yield about 71 g material containing about 99.1% reb B. The water solubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 4 Preparation of Reb B

75 g of material prepared according to EXAMPLE 2 was suspended in 1000 mL water. The mixture temperature was increased to 70° C. The suspension was held under moderate agitation conditions for 4 hours. The crystals were filtered and dried under vacuum to yield about 65 g material containing about 99.0% reb B. The water solubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 5 Preparation of Reb B Carboxylate Salt

100 g of reb B prepared according to EXAMPLE 4 was suspended in 1000 mL water. The suspension was held under moderate agitation conditions at room temperature for 1 hr. The pH of the mixture was adjusted to pH 11.0 with 1M KOH solution. The obtained mixture was agitated for 3 hrs. The crystals were filtered and washed with water on the filter until the washing water reached a neutral pH. The obtained crystalline material was dried under vacuum to yield about 95 g material. The water solubility (at 25° C.) of obtained material was about 0.1% (w/v).

Example 6 Preparation of Soluble Stevia Composition

50 g material prepared according to EXAMPLE 1 was mixed with 50 g of water and incubated in thermostated oil bath. The temperature was increased at 1° C. per minute to 121° C. The mixture was maintained at 121° C. for 1 hour and then the temperature was decreased to room temperature (25° C.) at 1° C. per minute. The solution was dried using YC-015 laboratory spray drier (Shanghai Pilotech Instrument & Equipment Co. Ltd., China) operating at 175° C. inlet and 100° C. outlet temperature. About 47 g of an amorphous powder was obtained with about 25% (w/v) solubility in water (at 25° C.).

Example 7 Preparation of Soluble Stevia Composition

42 g of reb A produced by PureCircle Sdn. Bhd. (Malaysia) with purity of 99.2% (dry basis) and 8 g of reb B prepared according to EXAMPLE 4 were mixed with 50 g of water and incubated in thermostated oil bath. The temperature was increased at 1° C. per minute to 121° C. The mixture was maintained at 121° C. for 1 hour and then the temperature was decreased to room temperature (25° C.) at 1° C. per minute. The solution was dried using YC-015 laboratory spray drier (Shanghai Pilotech Instrument & Equipment Co. Ltd., China) operating at 175° C. inlet and 100° C. outlet temperature. About 48 g of an amorphous powder was obtained with about 1.5% (w/v) solubility in water (at 25° C.).

Example 8 Preparation of Soluble Stevia Composition

42 g of reb A produced by PureCircle Sdn. Bhd. (Malaysia) with purity of 99.2% (dry basis) and 8 g of reb B potassium salt prepared according to EXAMPLE 5 were mixed with 50 g of water and incubated in thermostated oil bath. The temperature was increased at 1° C. per minute to 121° C. The mixture was maintained at 121° C. for 1 hour and then the temperature was decreased to room temperature (25° C.) at 1° C. per minute. The solution was dried using YC-015 laboratory spray drier (Shanghai Pilotech Instrument & Equipment Co. Ltd., China) operating at 175° C. inlet and 100° C. outlet temperature. About 49 g of an amorphous powder was obtained with about 2.5% (w/v) solubility in water (at 25° C.).

Example 9 Low-Calorie Orange Juice Drink

Orange concentrate (35%), citric acid (0.35%), ascorbic acid (0.05%), orange red color (0.01%), orange flavor (0.20%), and 0.05% stevia composition, were blended and dissolved completely in water (up to 100%) and pasteurized. The stevia composition was selected from a commercial stevia extract (containing stevioside 26%, rebaudioside A 55%, and 16% of other glycosides), a commercial rebaudioside A (containing 98.2% reb A) or material obtained according to EXAMPLE 7 and EXAMPLE 8.

The sensory evaluations of the samples are summarized in Table 1. The data show that the best results can be obtained by using the composition obtained according to EXAMPLE 8. Particularly the drinks prepared with said composition exhibited a rounded and complete flavor profile and mouthfeel.

TABLE 1 Evaluation of orange juice drink samples Comments Sample Flavor Aftertaste Mouthfeel Stevia Extract Sweet, licorice notes Bitterness and Not aftertaste acceptable Reb A Sweet, slight licorice Slight bitterness Not notes and aftertaste acceptable EXAMPLE 7 High quality sweetness, Clean, almost Full pleasant taste almost no bitterness similar to sucrose, and slight fairy rounded and aftertaste balanced flavor EXAMPLE 8 High quality sweetness, Clean, no Full pleasant taste similar bitterness and to sucrose, rounded no aftertaste and balanced flavor

The same method can be used to prepare juices and juice drinks from other fruits, such as apples, lemons, apricots, cherries, pineapples, mangoes, etc.

Example 10 Zero-Calorie Carbonated Beverage

Carbonated beverages made according to the formulas presented in Table 2 were prepared.

TABLE 2 Carbonated Beverage Formulas Quantity, % Stevia Ingredients Extract Reb A EXAMPLE 7 EXAMPLE 8 Cola flavor 0.340 0.340 0.340 0.340 ortho-Phosphoric 0.100 0.100 0.100 0.100 acid Sodium citrate 0.310 0.310 0.310 0.310 Sodium benzoate 0.018 0.018 0.018 0.018 Citric acid 0.018 0.018 0.018 0.018 Stevia composition 0.050 0.050 0.050 0.050 Carbonated water to 100 to 100 to 100 to 100

The sensory properties were evaluated by 20 panelists. The results are summarized in Table 3.

TABLE 3 Evaluation of zero-calorie carbonated beverage samples Number of panelists detected the attribute Stevia Taste attribute Extract Reb A EXAMPLE 7 EXAMPLE 8 Bitter taste 15 10 2 0 Astringent 16  9 1 0 taste Aftertaste 14 12 2 0 Comments Quality of Bitter Bitter Clean Clean sweet taste aftertaste aftertaste (17 of 20) (20 of 20) (15 of 20) (10 of 20) Overall Satisfactory Satisfactory Satisfactory Satisfactory evaluation (1 of 20) (5 of 20) (16 of 20) (20 of 20)

The above results show that the beverages prepared using the composition obtained according to EXAMPLE 8 possessed the best organoleptic characteristics.

Example 11 Diet Cookies

Flour (50.0%), margarine (30.0%) fructose (10.0%), maltitol (8.0%), whole milk (1.0%), salt (0.2%), baking powder (0.15%), vanillin (0.1%) and different stevia compositions (0.03%) were kneaded well in dough-mixing machine. The obtained dough was molded and baked in oven at 200° C. for 15 minutes. The stevia compositions were selected from a commercial stevia extract (containing stevioside 26%, rebaudioside A 55%, and 16% of other glycosides), a commercial rebaudioside A (containing 98.2% reb A) and material obtained according to obtained according to EXAMPLE 7 and EXAMPLE 8.

The sensory properties were evaluated by 20 panelists. The best results were obtained in samples containing the composition obtained according to EXAMPLE 8. The panelists noted a rounded and complete flavor profile and mouthfeel.

Example 12 Yoghurt

Different stevia compositions (0.03%) and sucrose (4%) were dissolved in low fat milk. The stevia compositions were selected from a commercial stevia extract (containing stevioside 26%, rebaudioside A 55%, and 16% of other glycosides), a commercial rebaudioside A (containing 98.2% reb A) and the material obtained according to EXAMPLE 7 and EXAMPLE 8. After pasteurizing at 82° C. for 20 minutes, the milk was cooled to 37° C. A starter culture (3%) was added and the mixture was incubated at 37° C. for 6 hours then at 5° C. for 12 hours.

The sensory properties were evaluated by 20 panelists. The best results were obtained in samples containing the composition obtained according to EXAMPLE 8. The panelists noted a rounded and complete flavor profile and mouthfeel.

It is to be understood that the foregoing descriptions and specific embodiments shown herein are merely illustrative of the best mode of the invention and the principles thereof, and that modifications and additions may be easily made by those skilled in the art without departing for the spirit and scope of the invention, which is therefore understood to be limited only by the scope of the appended claims. 

What is claimed is:
 1. A stevia composition comprising a carboxylate salt of rebaudioside B, made by a process comprising the steps of: providing a stevia sweetener; providing an alkaline aqueous solution; dispersing the stevia sweetener in the alkaline solution and incubating for about 12 to 48 hours at about 55-75° C. to form a mixture; cooling the mixture to about 10-30° C. and adjusting the pH with acid to about pH 3.0-4.0; incubating the mixture to obtain a precipitate; separating the precipitate and washing it with water; drying the washed precipitate to obtain the stevia composition; dispersing the stevia composition in water to form a mixture; adding a base corresponding to the carboxylate salt into the mixture to achieve a pH level of 6.5 to 14.0; incubating the mixture for 0.1 to 24 hours to facilitate conversion of a rebaudioside B into the carboxylate salt of rebaudioside B; and separating and drying the stevia composition comprising the carboxylate salt of rebaudioside B.
 2. The stevia composition of claim 1, the composition comprising a mixture of rebaudioside A, and rebaudioside B, and rebaudioside B carboxylate salt, wherein the ratio of rebaudioside A to rebaudioside B and rebaudioside B carboxylate salt ranges from about 5%:95% to about 95%:5% (w/w).
 3. The stevia composition of claim 1, the composition comprising a mixture of rebaudioside D, rebaudioside B, and rebaudioside B carboxylate salt, wherein the ratio of rebaudioside D to rebaudioside B and rebaudioside B carboxylate salt ranges from about 5%:95% to about 95%:5% (w/w).
 4. A food, beverage, pharmaceutical or cosmetic composition comprising the stevia composition of claim
 1. 5. The stevia composition of claim 1, wherein the process of making comprises before the step of dispersing the stevia composition in water to form a mixture the further steps of: suspending the stevia composition in water and incubating at an elevated temperature for about 1-3 hours, wherein the elevated temperature is a temperature of about 50 to 100° C.; separating the stevia composition from the water and drying the stevia composition to obtain a purified stevia composition; wherein the purified stevia composition comprises rebaudioside B with greater than about 99% purity.
 6. The stevia composition of claim 1, wherein the process of making comprises the further steps of: suspending the stevia composition comprising the carboxylate salt of rebaudioside B in water to form a suspension; increasing the temperature by a gradient heating method; holding the suspension at an elevated temperature, wherein the elevated temperature is a temperature of 110 to 140° C.; decreasing the temperature of the suspension to room temperature by a gradient cooling method to obtain a stevia composition solution; and spray drying the stevia composition solution to provide a stevia composition comprising the carboxylate salt of rebaudioside B with a solubility in water at 25° C. of about 2.5% (w/v).
 7. The stevia composition of claim 6, wherein the gradient of the gradient heating and/or gradient cooling method is 1° C. per minute. 